6. How to select a real-time fluorescent PCR instrument?
2026-03-24 11:18
(1) Popularity of use:
For non-new brands, to ensure sufficient reliability of the purchased instrument, PCR laboratories can judge by the popularity of the instrument at home and abroad. More widely used instruments are more verified by practice and thus more reliable;
(2)Detection throughput (number of reaction wells):
Select instruments with different throughputs according to laboratory requirements when purchasing quantitative PCR instruments. The detection throughput of real-time fluorescent PCR instruments on the market ranges from 16 to 384 wells. Laboratories can choose suitable instruments according to their testing projects and development status, and there is no need to pursue the most expensive ones. Generally, 96-well throughput is sufficient; adjust according to actual situations;
(3)Number of detection channels:
With the increasing number of projects (e.g., gene expression, single nucleotide polymorphism analysis, high-resolution melting curve analysis), single-channel instruments cannot meet laboratory needs, while multi-channel design facilitates the detection of multiplex PCR and its derivative analysis modes such as gene expression;
(4)Consumable openness:
The openness of consumables such as amplification reaction tubes may determine the level of daily testing costs. PCR laboratories can consider this when selecting real-time fluorescent PCR instruments. However, RNase-free consumables should be used for projects detecting RNA as pathogen genetic material;
(5) Hardware design features:
Real-time fluorescent PCR instruments are mainly divided into 96-well plate type and centrifugal type, each with unique advantages and inevitable defects. 96-well plate instruments can hold a large sample volume (up to 100ul) without special consumables. Traditional 96-well plate instruments mostly adopt halogen lamp excitation and CCD detection, with advantages of higher light intensity and wider wavelength range than LED lamps (excellent for fluorescent dye excitation, stable performance, lifespan of about 3000 hours), but CCD detection may cause edge effects due to different optical paths from each sample well to the light source and detector, affecting results to a certain extent. Centrifugal instruments usually adopt LED excitation and PMT detection, avoiding edge effects in design. PMT detection can amplify or reduce fluorescent signals, being more flexible and sensitive than CCD detection, but LED has lower fluorescence intensity and narrower wavelength range, resulting in poorer excitation effect on fluorescent dyes than halogen lamps;
(6)Running speed:
Among the many technical parameters of real-time fluorescent PCR instruments, the temperature rise and fall speed is also very important. Faster temperature rise and fall can shorten reaction time, reduce possible non-specific binding and reaction time, and improve PCR specificity. However, faster running speed has higher requirements for the heating and cooling module, so stability is the biggest problem. For instruments with long-term market verification and good stability, faster running speed brings more convenience;
(7)Flexibility:
On the premise of guaranteed basic performance, consider the instrument flexibility, mainly including transportation flexibility, disassembly flexibility, software upgrade flexibility, module replacement flexibility and operation flexibility.
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